目的 探讨水飞蓟宾对H₂O₂诱导状态下H9C2心肌细胞氧化应激损伤的保护作用。方法 将大鼠H9C2心肌细胞分为对照组、H₂O₂(200 μmol/L)干预组以及水飞蓟宾(100、200 μmol/L)+H₂O₂(200 μmol/L)干预组,每组设6个复孔。各组经过药物干预6 h后,通过Giemsa染色法观察细胞形态学,通过MTT法测定细胞存活率、流式细胞术测定细胞凋亡率;测定细胞培养液中乳酸脱氢酶(LDH)、磷酸激酶(CK)、谷草转氨酶(AST)活性和丙二醛(MDA)含量;测定细胞中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性。结果 与对照组比较,H₂O₂干预组H9C2心肌细胞形态明显异常、存活率显著降低且凋亡率显著升高,培养液中LDH、CK、AST活性和MDA含量均显著升高,细胞中SOD、CAT、GSH-Px活性显著降低,差异均具有统计学意义(P < 0.05)。与H₂O₂干预组比较,水飞蓟宾(100、200 μmol/L)+H₂O₂(200 μmol/L)干预组培养液中AST、CK活性和MDA含量均显著降低,差异具有统计学意义(P < 0.05);水飞蓟宾(200 μmol/L)+H₂O₂(200 μmol/L)干预组H9C2心肌细胞形态明显改善、存活率显著升高、凋亡率显著降低,培养液中LDH活性显著降低,细胞中SOD、CAT、GSH-Px活性显著升高,差异均具有统计学意义(P < 0.05)。结论 水飞蓟宾能够有效改善H₂O₂诱导状态下H9C2心肌细胞形态,提高其存活率并降低凋亡率,改善细胞中抗氧化酶活性、降低细胞损伤,提示水飞蓟宾对H₂O₂诱导H9C2心肌细胞氧化应激损伤具有剂量相关性的保护作用。

Objective To investigate the protective effects of silibinin on oxidative stress injury of H9C2 cardiomyocytes induced by H₂O₂. Methods H9C2 cardiomyocytes were randomly divided into control group, H₂O₂ (200 μmol/L) group, silibinin (100 μmol/L) +H₂O₂ (200 μmol/L) group, and silibinin (200 μmol/L) +H₂O₂ (200 μmol/L) group, and each group set six holes. After H₂O₂ stimulation for 6 h, the morphology changes were observed by microscope, and the survival rate and the apoptosis rate were detected by MTT method. The activity of LDH, CK, & AST, and the content of MDA in culture medium were detected, and the activities of SOD, CAT, and GSH-Px in cardiomyocytes were also determined. Results Compared with the control group, the morphology of H9C2 cardiomyocytes in H₂O₂ (200 μmol/L) group was abnormal, the survival rate was significantly decreased, and the apoptosis rate was significantly increased (P < 0.05). The activity of LDH, CK, & AST, and the content of MDA in culture medium were significantly increased (P < 0.05), the activity of SOD, CAT, and GSH-Px in cardiomyocytes were significantly decreased (P < 0.05). Compared with the H₂O₂ group, the activity of CK, AST, and the content of MDA in culture medium of silibinin (100, 200 μmol/L) +H₂O₂ (200 μmol/L) groups were significantly decreased (P < 0.05). The morphology of H9C2 cardiomyocytes in silibinin (200 μmol/L) +H₂O₂ (200 μmol/L) group was improved, the survival rate was significantly increased, and the apoptosis rate was significantly decreased (P < 0.05). The activity of LDH was significantly decreased (P < 0.05), and the activity of SOD, CAT, and GSH-Px in cardiomyocytes were significantly increased (P < 0.05). Conclusion Silibinin can effectively improve the morphology of H9C2 cardiomyocytes induced by H₂O₂, increase the survival rate and decrease the apoptosis rate, improve the activity of antioxidase, and depress the cell injury, which suggests that silibinin has dose-dependent protective effects against the oxidative stress of H9C2 cardiomyocytes induced by H₂O₂.

河北省卫生厅重点科技研究计划项目(20130359)